RU2656228C9 - WEAKLY CRYSTALLISED β-MODIFICATION OF (S)-ISOPROPYL 2-((S)-(((2R,3R,4R,5R)-5-(2,4-DIOXO-3,4-DIHYDROPYRIMIDIN-(2H)-YL)-4-FLUORO-3-HYDROXY-4-METHYLTETRAHYDROFURAN-2-YL)METHOXY)-(PHENOXY)PHOSPHORYLAMINO)PROPANOATE, METHOD FOR PRODUCTION THEREOF AND PHARMACEUTICAL COMPOSITION BASED THEREON - Google Patents

Abstract

FIELD: chemistry.SUBSTANCE: invention relates to a new anhydrous modification of (S)-isopropyl 2-((S)-(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-(phenoxy)phosphorylamino)propanoate for the treatment of chronic viral hepatitis C, a method for its preparation and a pharmaceutical composition based thereon, which can be used in the pharmaceutical industry. Disclosed is a new β-modification of (S)-isopropyl 2-((S)-(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-(phenoxy)phosphorylamino)propanoate, characterised by the following set of interplanar distances (d, Å) and the corresponding intensities (I, %): 25.638 Å – 15.95 %; 21.101 Å – 17.80 %; 16.458 Å – 21.57 %; 13.368 Å – 26.61 %; 12.290 Å – 28.21 %; 11.343 Å – 31.14 %; 11.032 Å – 40.14 %; 10.287 Å – 36.18 %; 9.083 Å – 39.89 %; 8.458 Å – 43.59 %; 7.641 Å – 40.40 %; 7.148 Å – 46.78 %; 7.013 Å – 54.75 %; 6.528 Å – 46.01 %; 5.975 Å – 55.33 %; 5.225 Å – 79.00 %; 5.093 Å – 100.00 %; 4.892 Å – 86.15 %; 4.707 Å – 98.40 %; 4.614 Å – 84.56 %; 4.535 Å – 94.38 %; 4.405 Å – 90.68 %; 4.371 Å – 89.60 %; 4.259 Å – 75.49 %; 4.008 Å – 62.48 %; 3.792 Å – 57.69 %; 3.741 Å – 54.50 %; 3.539 Å – 47.35 %; 3.500 Å – 46.01 %; 3.263 Å – 46.27 %; 3.172 Å – 38.54 %; 3.141 Å – 40.71 %; 2.999 Å – 25.82 %; 2.839 Å – 25.82 %; 2.773 Å – 25.06 %; 2.722 Å – 27.38 %; 2.542 Å – 25.02 %; 2.359 Å – 23.68 %; 2.192 Å – 22.85 %; 2.117 Å – 21.83 %, and the presence of two endothermic effects on the DSC curve at temperatures 39.7–48.1 °C and 120.7–125.9 °C with improved efficiency.EFFECT: disclosed is a new effective method for the preparation of said form, characterised in that sofosbuvir is dissolved in an organic solvent at 25–70 °C, the resulting solution is mixed with distilled water in a ratio of 1:2–1:10, frozen at a cooling rate of at least 60 deg/min, freeze-dried and further annealed at temperatures of 50–85 °C for at least 1 hour, followed by cooling to room temperature at a rate not higher than 5 deg/h.4 cl, 4 tbl, 6 dwg, 5 ex

Description

The invention relates to organic chemistry and relates to a new slightly crystallized modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- ( 2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate [sofosbuvir (international unlicensed name)], which we called the weakly crystallized β-modification, its preparation and pharmaceutical composition based on it, which can be used in the pharmaceutical industry and medicine as a nucleotide inhibitor olimerazy to treat chronic hepatitis C.

The term “slightly crystallized” was introduced in the name of the new modification due to the diffraction pattern of the obtained sofosbuvir modification that contains broadened diffraction maxima, unlike radiographs of known crystal modifications, where the peaks are well pronounced or there is a X-ray amorphous halo without maxima in the case of amorphous modification. If the powder obtained by us were amorphous, then there would be no distinct peaks on the thermoanalytical curves. It would be the displacement of the baseline of the DSC - curves associated with the glass transition [http://www.pslc.ws/english/dsc.htm].

Known derivatives of nucleoside phosphoroamidates and their pharmaceutically acceptable salts as a nucleotide polymerase inhibitor, methods for their preparation and pharmaceutical compositions containing them for the treatment of chronic viral hepatitis C. (RU 2478104 C2 of 03/27/2013; RU 2009139968 A of May 10, 2011; WO 2008/121634 A2 dated October 9, 2008; US 7429572 B2 dated September 25, 2008, etc.).

However, in these patents, the synthesized substances are not characterized as to their belonging to one or another polymorphic modification.

The crystalline and amorphous modifications of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) are known -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate as solvates, as well as salts of sofosbuvir with pharmaceutically acceptable acids. (EA 2011/71417 A1 dated 05/25/2012; WO 2011/123645 A2 dated 10/6/2011; WO 2016/023905 A1 dated 02/18/2016, etc.)

However, all these forms are different from the proposed new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxco3, 4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate by chemical composition.

Closest to the claimed new, not previously known, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4 -dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate are the polymorphic modifications of sofosbuvir: crystalline forms: Form 1; Form 6; Form 7; Form 8; Form A; Form C; Form D; Form E; Form M; Form M1; Form M; Form Z-1; the crystalline form and the amorphous form (WO 2010/135569 A1 dated November 25, 2010; WO 2015/025853 A1 dated March 5, 2015; WO 2015/097605 A1 dated April 2, 2015; WO 2015/099989 A1 from 2.07. 2015; CN 104125252 A dated November 5, 2014; WO 2016/035006 A1 dated March 10, 2016; WO 2015/126995 A1 dated August 27, 2015; WO 2015/191945 A2 dated December 17, 2015; WO 2016/038542 A2 dated March 17, 2016; WO 2016/008461 A1 dated January 21, 2016, CN 104277088 A dated January 14, 2015; WO 2016/023906 A1 February 18, 2016, etc.).

Known crystalline modifications of sofosbuvir are characterized by physicochemical analysis methods such as: nuclear magnetic resonance (NMR) - a combination of chemical shifts (ppm), high performance liquid chromatography (HPLC) - retention times (min), x-ray phase analysis (XRF) - sets of angles (2θ, degrees), interplanar distances (d, Å) and their intensity (I, imp./min; I _rel , = I _i / I _max × 100,%) or directly by X-ray diffraction (the diffractograms obtained by the authors of the above patents on Cu Kα - radiation), term analytical studies (temperatures of the thermal effects), and others.

The similarity of the proposed new, not previously known, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin - (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, with known crystalline and amorphous modifications of sofosbuvir, is the identity of their chemical compositions.

The difference between the claimed new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine - (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, from the known crystalline and amorphous modifications of sofosbuvir lies in the difference of their X-ray diffraction analysis of X-ray diffraction (XRF) ): a certain set of angles 2θ (deg) - interplanar distances (d, Å), and their intensity (I _rel , = I _i / I _max × 100,%), as well as in the difference between the curves of the differential scanning curve Rimetry (DSC). The DSC curve of a new weakly crystallized β-modification of sofosbuvir is characterized by two endothermic effects.

Known crystalline modifications of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4 -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate are white or almost white powders, odorless, easily soluble in acetone and ethanol, soluble in isopropanol, little soluble in water.

The sofosbuvir modifications discussed above are obtained from the starting components or one from the other in solutions, varying the type of solvents, the conditions for preparing solutions, and the conditions for isolating the target product from organic solvents or their mixtures by various methods, such as crystallization from solutions, followed by filtration, washing and further, by air drying, vacuum drying, spray drying or freeze drying (WO 2010/135569 A1 dated November 25, 2010; WO 2015/025853 A1 dated March 5, 2015; WO 2015/099989 A1 from 2.07.2015 g .; CN 104125252 A dated November 5, 2014; WO 2016/035006 A1 dated March 10, 2016; WO 2015/126995 A1 dated August 27, 2015; WO 2015/191945 A2 dated December 17, 2015; WO 2016/038542 A2 dated March 17, 2016; WO 2016 / 008461 A1 dated 01/21/2016, CN 104277088 A dated 01/14/2015, WO 2016/023905 A1 February 18, 2016; WO 2016/023906 A1 dated February 18, 2016, etc.).

The closest in technical essence is a method for producing an amorphous form of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- ( 2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, consisting in the fact that sofosbuvir is dissolved in a mixture of water and an organic solvent, using ultrasound to accelerate dissolving, homogeneous solution is frozen in a bath with liquid nitrogen and lyophilized (subjected to freeze-drying) at -36 ° C and pressure from 0 to 2 mbar. The rate of freezing of a homogeneous solution and the duration of lyophilization were not indicated by the authors of the patent (WO 2016/023905 A1 dated February 18, 2016).

The difference in the method of obtaining the claimed new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4 -dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate from a process for the preparation of the known crystalline forms of sofosbuvir is that sofosbuvir is dissolved in organic solvent at 25 ° C-70 ° C, mix the resulting solution with distilled water in a ratio of 1: 2 - 1:10, freeze at a cooling rate not lower than 60 deg / min, subjected to by freeze-drying and further annealed at temperatures (50-85) ° C for at least 1 hour, followed by cooling to room temperature at a rate no higher than 5 ° / hr.

The difference of the claimed pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C based on (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo- 3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate from known compositions (Instructions for use of the drug sofosbuvir: Sovaldi ^® / Sovaldi ^® , Viropack ^® / MPI Viropack ^® , Hepcinat ^® , Hopetavir ^® , etc.) is that a therapeutically effective amount of a new one is used as the active substance, not previously known, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate (sofosbuvir).

The aim of the invention is to obtain a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine - (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, develop a method for its preparation and use it in a pharmaceutical composition as a nucleotide polymerase inhibitor for treatment of chronic viral hepatitis C.

The goal has been achieved by the present invention, namely, obtaining a new, not previously known, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2.4- Dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, characterized by the following set of interplanar distances (d, Å ) and their corresponding intensities (I _from. , = I _max × 100,%):

25.638 Å - 15.95%; 21.101 Å - 17.80%; 16.458 Å - 21.57%; 13.368 Å - 26.61%; 12.290 Å - 28.21%; 11.343 Å - 31.14%; 11.032 Å - 40.14%; 01.287 Å - 36.18%; 9.083 Å - 39.89%; 8.458 Å - 43.59%; 7.641 Å - 40.40%; 7.148 Å - 46.78%; 7.013 Å - 54.75%; 6.528 Å - 46.01%; 5.975 Å - 55.33%; 5.225 Å - 79.00%; 5.093 Å - 100.00%; 4.892 Å - 86.15%; 4.707 Å - 98.40%; 4.614 Å - 84.56%; 4.535 Å - 94.38%; 4.505 Å - 90.68%; 4.371 Å - 89.60%; 4.259 Å - 75.49%; 4.008 Å - 62.48%; 3.792 Å - 57.69%; 3.741 Å - 54.50%; 3.539 Å - 47.35%; 3,500 Å - 46.01%; 3.263 Å - 46.27%; 3.172 Å - 38.54%; 3.141 Å - 40.71%; 2.99 Å - 25.82%; 2.839 Å - 25.82%; 2.773 Å - 25.06%; 2.722 Å - 27.38%; 2.522 Å - 25.02%; 2.359 Å - 23.68%; 2.192 Å - 22.85%; 2.117 Å - 21.83%,

and the presence of two endothermic effects on the DSC curve at temperatures (39.7-48.1) ° C and (120.7-125.9) ° C.

The goal was also achieved by developing a method for obtaining a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo- 3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, which consists in that sofosbuvir is dissolved in an organic solvent at 25 ° C-70 ° C, mix the resulting solution with distilled water in a ratio of 1: 2 - 1:10, freeze at a cooling rate not lower than 60 deg / min, freeze-dry and add tively annealed at temperatures (50-85) ° C for at least 1 hour, followed by cooling to room temperature at a rate no higher than 5 ° / hr.

In addition, the goal was achieved by applying a new, not previously known, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo- 3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate for preparing a pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C. The composition indicated contains weakly crystallized β-modification of sofosbuvir in a therapeutically effective amount and, at least one pharmaceutically acceptable carrier, diluent, excipient, or vehicle.

From the patent and scientific literature is not known weakly crystallized β - modification of N (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4 -dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, its preparation and its use in the pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C.

We have discovered a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine - (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, characterized by a specific set of diffraction maxima (d, Å) and their intensity (I _rel. ,%) and the presence of two endothermic effects on the DSC curve, a method for its preparation and its use for the preparation of a pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of I have chronic viral hepatitis C.

The inventive new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is a white or almost white powder, odorless, easily soluble in acetone and ethanol , soluble in isopropanol, little soluble in water.

To clarify the essence of the proposed technical solution, the following tables and figures are attached to the description:

Table 1. Angles 2θ, hail, interplanar distances (d, Å) and their intensities (I _rel ,%) of the known modifications of sofosbuvir according to the literature data.

Table 2. Angles 2θ, hail, interplanar distances (d, Å) and their intensities (I _rel ,%) of the known crystalline modification of sofosbuvir (Form I), which was used as the starting material.

Table 3. Angles 2θ, interplanar distances (d, Å) and their intensities (I _rel ,%) of a new, previously unknown, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - ((2R, 3R , 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Table 4. The results of thermoanalytical studies of known modifications of sofosbuvir according to literature data.

Fig. 1. NMR ¹ H - the spectrum of the known crystalline modification of sofosbuvir (Form I), which was used as the starting material.

Fig. 2. NMR ¹ H - spectrum of a new, not previously known, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo- 3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Fig. 3. Diffractogram of a known crystalline modification of sofosbuvir (Form I), which was used as the starting material.

Fig. 4. A typical diffractogram of a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4 -dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Fig. 5. Thermogram of the known crystalline modification of sofosbuvir (Form 1), which was used as the starting material.

Fig. 6. Typical thermogram of a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4 -dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

To identify the obtained substance, a complex of physicochemical methods of analysis was carried out.

Initially, nuclear magnetic resonance (NMR ¹ H) and high performance liquid chromatography (HPLC) were used to establish the identity of the chemical formulas of the known crystalline modification of sofosbuvir (Form 1), which was used as the starting material, and the new, previously unknown, weakly crystallized β - modification (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro- 3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

The chemical shifts of the starting sofosbuvir (Form 1) and the substance obtained from it were carried out in its saturated solution in deuterated dimethyl sulfoxide (DMSO-D ₆ ) on a VXR-400 high-resolution NMR spectrometer from the United States. The data obtained are shown in Fig. 1 and 2, respectively. Comparison of the results presented in Fig. 1 [known crystalline modification of sofosbuvir (Form 1)], with the data shown in fig. 2 [a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine - (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate shows that the NMR - ¹ H - spectra of the starting and the substances we obtained are almost identical, those. the resulting material is (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4 -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Chromatographic mobility of a known crystalline modification of sofosbuvir (Form 1), which was used as a starting material and, obtained from it, a new, not previously known, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy ) phosphorylamino) propanoate was determined by HPLC on a Shimadzu LC-20A chromatograph with an SPD-M20A detector and with a column 150 mm long, 4.6 mm in diameter, XSelect HSS Т3, 3.5 µm sorbent with detection at d the wavelength of 260.8 nm. A 0.05% solution of orthophosphoric acid in water was used as the mobile phase A, acetonitrile was used as the mobile phase B, and an acetate buffer solution with pH = 4: methanol = 60: 40 at 25 ° C and a rate of 1.3 ml was used as a sample solvent. / min The volume of the test sample was 5 μl with a concentration of 1.0 mg / ml. It turned out that the chromatograms of the known crystalline modification of sofosbuvir (Form 1), which was used as a starting material and a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - ((2R, 3R, 4R , 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate contains one peak of the main component with characteristic retention times of 16.187 minutes and 16.167 minutes, respectively. These values of retention times, within the error of determination - 0.06%, are the same. The purity of the investigated samples, determined by HPLC, was 100.00% (starting material) and 99.97% (new, not previously known, slightly crystallized β - modification of sofosbuvir).

Experiments carried out by the HPLC method indicate that upon receipt of a new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2 , 4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate from the starting material - known crystalline modification sofosbuvir (Form 1), decomposition of the original substance does not occur.

Thus, the experimental results of nuclear magnetic resonance (NMR- ¹ H) and high-performance liquid chromatography (HPLC) clearly indicate that the substance obtained is (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R , 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

To confirm that the resulting substance is new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxco3 , 4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, X-ray phase analysis (XRPA) and thermoanalytical studies were carried out.

X-ray phase analysis (XRD) of a known crystalline modification of sofosbuvir (Form 1), which was used as the starting material and the compound obtained from it was carried out on a Rigaku D / MAX-2500 diffractometer (Rigaku, Japan) using CuKα radiation (λ = 1.54056 Å) .

The obtained X-ray diffraction data for sofosbuvir, which was used as the starting material, is given in Table. 2 and fig. 3. Comparison of the results of XRF, presented in table. 2 and in table. 1 (literature data), clearly indicates that the known crystalline modification of sofosbuvir was used as the starting material - Form 1. The results of X-ray phase analysis of the powder obtained from the known crystalline modification of sofosbuvir (Form 1) are presented in Table. 3 and fig. 4. Comparison of the data presented in table. 3, tab. 2 and tab. 1, and also on fig. 3, Fig. 4 and in the figures cited in the literature (WO 2010/135569 A1 dated November 25, 2010; WO 2015/025853 A1 dated March 5, 2015; WO 2015/097605 A1 dated July 2, 2015; WO 2015/099989 A1 from 07.27.2015, CN 104125252 A from 5.11.2014, WO 2016/035006 A1 from 03.10.2016, WO 2015/126995 A1 from 08.27.2015, WO 2015/191945 A2 from 17.12. 2015; WO 2016/038542 A2 dated 03/17/2016; WO 2016/008461 A1 dated 01/21/2016; CN 104277088 A dated January 14, 2015; WO 2016/023906 A1 dated February 18, 2016, etc. .) suggests that the resulting sofosbuvir powder is a new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2, 4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahyd irofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Thus, the experimental results of X-ray phase analysis unequivocally indicate that the resulting substance is new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) - 5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. It is characterized by a set of interplanar distances (d, Å) and corresponding intensities (I _rel ,%), different from other crystalline modifications of sofosbuvir:

25.638 Å - 15.95%; 21.101 Å - 17.80%; 16.458 Å - 21.57%; 13.368 Å - 26.61%; 12.290 Å - 28.21%; 11.343 Å - 31.14%; 11.032 Å - 40.14%; 01.287 Å - 36.18%; 9.083 Å - 39.89%; 8.458 Å - 43.59%; 7.641 Å - 40.40%; 7.148 Å - 46.78%; 7.013 Å - 54.75%; 6.528 Å - 46.01%; 5.975 Å - 55.33%; 5.225 Å - 79.00%; 5.093 Å - 100.00%; 4.892 Å - 86.15%; 4.707 Å - 98.40%; 4.614 Å - 84.56%; 4.535 Å - 94.38%; 4.505 Å - 90.68%; 4.371 Å - 89.60%; 4.259 Å - 75.49%; 4.008 Å - 62.48%; 3.792 Å - 57.69%; 3.741 Å - 54.50%; 3.539 Å - 47.35%; 3,500 Å - 46.01%; 3.263 Å - 46.27%; 3.172 Å - 38.54%; 3.141 Å - 40.71%; 2.99 Å - 25.82%; 2.839 Å - 25.82%; 2.773 Å - 25.06%; 2.722 Å - 27.38%; 2.522 Å - 25.02%; 2.359 Å - 23.68%; 2.192 Å - 22.85%; 2.117 Å - 21.83%.

Thermoanalytical studies of the known crystalline modification of sofosbuvir (Form 1), which was used as a starting material and the compound obtained from it, was carried out on a DSC 204 F1 Peu-nix (NETZSCH, Germany) in an argon flow with increasing temperature at a rate of 10 deg / min. Aluminum cuvettes were used as sample holders. The sample weight was (5-10) mg. The data obtained are shown in Fig. 5 and fig. 6. It can be seen that on the curve of the differential scanning calorimetry of a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - ((2R, 3R, 4R, 5R) -5- (2.4 -dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate (Fig. 6) 2 endothermic peaks are observed at temperatures (39.7-48.1) ° C and (120.7-125.9) ° C, and on the DSC curve of the known sofosbuvir crystal modification Form 1 there is only one endothermic peak at (97.4-106, 7) ° С (see fig. 5). Note that according to the literary sources presented in table. 4, on thermograms of known crystal modifications, there is one endothermic peak.

Thus, the conducted thermoanalytical experiments showed that the obtained new, not previously known, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2.4 -dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is characterized by the presence of two endothermic effects on the DSC curve at temperatures (39.7-48.1) ° С and (120.7-125.9) ° С unlike the known modifications of sofosbuvir.

The experimental data of X-ray phase analysis and thermoanalytical studies show unequivocally that the newly obtained, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - ((2R, 3R, 4R, 5R) -5 - (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is characterized by a different crystalline modifications of sofosbuvir with a set of interplanar distances and corresponding intensities and the presence of two endo thermal effects on the DSC curve.

The method of obtaining a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine - (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is that sofosbuvir is dissolved in an organic solvent at 25 ° C-70 ° C, mix the resulting solution with distilled water in a ratio of 1: 2 - 1:10, freeze at a cooling rate of at least 60 degrees / min, freeze-drying and additionally anneal at temperatures (50-85) ° C for Not less than 1 hour, followed by cooling to room temperature at a rate not exceeding 5 deg / h.

The difference in the method of obtaining the proposed new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxco3,4-dihydropyrimidine - (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate from the method of obtaining the known crystalline forms of sofosbuvir is that sofosbuvir is dissolved in an organic solvent at 25 ° C-70 ° C, mix the resulting solution with distilled water in a ratio of 1: 2 - 1:10, freeze at a cooling rate not lower than 60 deg / min, subject to Blima-drying and additionally annealed at temperatures (50-85) ° C for at least 1 hour, followed by cooling to room temperature at a rate not exceeding 5 deg / h.

The proposed method of obtaining, consisting in the fact that sofosbuvir is dissolved in an organic solvent at 25 ° C-70 ° C, the resulting solution is mixed with distilled water in a ratio of 1: 2 - 1:10, frozen at a cooling rate of at least 60 degrees / min, subjected to freeze-drying and further annealed at temperatures (50-85) ° C for at least 1 hour, followed by cooling to room temperature at a rate not exceeding 5 deg / h, allows you to get a new, not previously known, slightly crystallized β-modification (S ) -isopropyl 2- ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran -2-yl) methoxy) - (phenoxy) phosphoryl-amino) propanoate.

As an organic solvent, it is possible to use lower alcohols, acetone, acetonitrile, dimethyl sulfoxide, and others that are miscible with water.

The preparation of the sofosbuvir solution at elevated temperatures was used to accelerate the dissolution process of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxco3,4-dihydro-pyrimidine- ( 2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

Reducing the temperature of the sofosbuvir solution in an organic solvent below 25 ° C (room temperature) is not advisable because it requires additional energy costs for cooling the system.

Increasing the temperature of the sofosbuvir solution in an organic solvent above 70 ° C is also not advisable because of the need to use special equipment.

Mixing an organic solution of sofosbuvir in an organic solvent with water in a ratio of less than 1: 2 leads to the fact that under these conditions one of the known modifications of sofosbuvir is formed. Obtain a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails.

Mixing an organic solution of sofosbuvir in an organic solvent with water in a ratio of more than 1:10 does not lead to an increase in the yield of the target product — a new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy ) phosphorylamino) propanoate and only increases the cost of production.

The decrease in the freezing rate below 60 deg / min, the cryocrystallization rate of the sofosbuvir solution decreases and approaches the equilibrium process. This results in already known modifications. Obtain a new, previously unknown, crystalline β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails. Note that the known method of freezing a solution containing sofosbuvir (WO 2016/023905 A1 02.18.2016) suggests freezing in a bath with liquid nitrogen. Under these conditions, as a rule, the freezing rate is not higher than 50 deg / min.

Freeze-drying of a frozen solution of sofosbuvir is necessary to obtain a dry powder, which further allows to form a new, actually unknown, slightly-crystallized β-modification of (S) -isopropyl 2 - ((S) - ((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. The preferred mode of freeze-drying is a process that is carried out at temperatures: on the condenser -42 ° C ... -56 ° C; on the product -196 ... + 50 ° C and residual pressure in the chamber (9-3) × 10 ^-2 Torr for 22-26 hours

Additional annealing of freeze-drying products at temperatures of (50-85) ° C for at least 1 hour, followed by cooling to room temperature at a rate not exceeding 5 deg / h, allows to obtain a new, not previously known, slightly crystallized β-modification (S ) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3- hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate.

The decrease in the temperature of additional annealing of products of freeze-drying below 50 ° C leads to the fact that under these conditions one of the known modifications of sofosbuvir is formed. Obtain a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo3,4-dihydro-pyrimidine- (2H ) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails.

An increase in the temperature of additional annealing of freeze-drying products above 85 ° C leads to the fact that under these conditions one of the known modifications of sofosbuvir is formed. Obtain a new, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo3,4-dihydro-pyrimidine- (2H ) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails.

Exposure of products of freeze-drying at temperatures (50-85) ° C for less than 1 hour leads to the fact that under these conditions one of the known modifications of sofosbuvir is formed. Obtain a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails.

Increasing the cooling rate of the powder from a temperature of (50-85) ° C to room temperature at a speed higher than 5 degrees / hour results in one of the known modifications of sofosbuvir being formed under these conditions. Obtain a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in pure form fails.

The possibility of carrying out the invention is illustrated by the following examples, but is not limited to them.

Example 1. In 35 ml of an organic solvent - methyl alcohol brand "HCH" GOST 2222-95 of the company "LLC Vekton-M, Russia" with stirring, dissolve 0.35 g of the known crystalline modification of sofosbuvir (Form 1) at 25 ° C, mix the resulting a solution with 70 ml of distilled water (1: 2 ratio) and freeze at a rate of 60 deg / min by pouring the entire volume of the solution into a pre-filled with liquid nitrogen (T _Kip = -196 ° C), but not previously cooled, stainless steel tray. The frozen product on the pallet is transferred to a sublimation chamber and freeze-dried at temperatures: on a condenser of -52 ° C ... -56 ° C; on the product -196 ° C ... + 30 ° C and a residual pressure in the chamber (9-5) 10 ^-2 Torr for 26 hours. The product of freeze-drying is a light, fluffy white powder, placed in a heat-resistant chemical glass with a capacity of 100 ml , bring it into a drying cabinet, preheated to 50 ° C, kept at this temperature for 1 hour and cooled to room temperature within 12.5 hours (cooling rate - 2 deg / h). The product yield was 0.34 g (97.1 wt.%). According to XRD data, the obtained substance is characterized by a typical diffractogram shown in Fig. 4 and a set of interplanar distances and their intensities that coincide with the corresponding values for the new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is presented in Table. 3. According to thermoanalytical studies, the resulting substance is characterized by a typical differential scanning calorimetry curve presented in Fig. 6, which characterizes a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. The temperatures of the endothermic peaks of the obtained powder are (39.7-48.1) ° C and (120.7-125.9) ° C. According to HPLC, the purity of the obtained new, previously unknown, weakly crystallized β-modification of sofosbuvir is 99.98%.

Example 2. In 60 ml of an organic solvent — acetonitrile, grade “ХЧ”, Specification 6-094326-76, produced by LLC Component - Reagent, RF, 0.90 g of the known crystalline modification of sofosbuvir (Form 1) is dissolved at 70 ° C with stirring , mix the resulting solution with 300 ml of distilled water (1: 5 ratio) and freeze at a rate of (2-3) × 10 ² deg / min by injecting the solution in small portions into a pre-cooled and filled with liquid nitrogen (T _Kip = -196 ° C) stainless steel tray. The frozen product on the pallet is transferred to a freeze-drying chamber and freeze-dried at temperatures: on a condenser of -45 ° C ... -51 ° C; on the product - 196 ° C ... + 50 ° C and a residual pressure in the chamber (9-5) 10 ^-2 Torr for 22 hours. The product of freeze-drying is a light, fluffy white powder, placed in a heat-resistant chemical glass with a capacity of 250 ml , bring it into a drying cabinet, preheated to 85 ° C, kept at this temperature for 2 hours and cooled to room temperature for 12.0 hours (cooling rate - 5 degrees / hour). The product yield was 0.87 g (96.7 wt.%). According to XRD data, the obtained substance is characterized by a typical diffractogram shown in Fig. 4 and a set of interplanar distances and their intensities that coincide with the corresponding values for the new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is presented in Table. 3. According to thermoanalytical studies, the resulting substance is characterized by a typical differential scanning calorimetry curve presented in Fig. 6, which characterizes a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. The temperatures of the endothermic peaks of the obtained powder are (37.5-49.3) ° C and (118.4-124.2) ° C. According to HPLC, the purity of the new, previously unknown, weakly crystallized β-modification of sofosbuvir is 99.96%.

Example 3. 40 ml of an organic solvent - dimethyl sulfoxide (DMSO), mark “ХЧ” TU 6-09-3818-77 produced by OOO NPP Viking, RF, with stirring dissolve 0.50 g of a known crystalline modification of sofosbuvir (Form 1) at 60 ° C, mix the resulting solution with 400 ml of distilled water (1:10 ratio) and freeze at a rate of about 10 ³ deg / min by spraying the solution with a pneumatic nozzle at a pressure of 0.4 kg / cm ² into a reactor filled with liquid nitrogen ( T _kip = -196 ° C). The frozen product is transferred to a stainless steel tray and subjected to freeze-drying at temperatures: on a -52 ° C… -56 ° C condenser; on the product -196 ° C ... + 40 ° C, and the residual pressure in the chamber (6-3) ⋅10 ^-2 Torr for 24 hours. The product of freeze-drying is a light, fluffy white powder, placed in a heat-resistant chemical glass with a capacity of 250 ml, bring it into a drying cabinet preheated to 70 ° C, kept at this temperature for 3 hours and cooled to room temperature for 15.0 hours (cooling rate - 3.0 degrees / hour). The product yield was 0.47 g (94.0 wt.%). According to XRD data, the obtained substance is characterized by a typical diffractogram shown in Fig. 4 and a set of interplanar distances and their intensities that coincide with the corresponding values for the new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is presented in Table. 3. According to thermoanalytical studies, the resulting substance is characterized by a typical differential scanning calorimetry curve presented in Fig. 6, which characterizes a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. The temperatures of the endothermic peaks of the obtained powder are (40.8-46.9) ° C and (121.8-127.1) ° C. According to HPLC, the purity of the new, previously unknown, weakly crystallized β-modification of sofosbuvir is 99.97%.

Example 4. In 200 ml of an organic solvent - ethyl alcohol of 95% GOST 51652 - 2000, produced by LLC Spirttorg, RF, 15.00 g of known crystalline modification of sofosbuvir (Form 1) are dissolved with stirring at 40 ° C, the resulting solution is mixed with 1800 ml distilled water (1: 9 ratio) and frozen at a rate of (2-3) × 10 ² deg / min by pouring the solution in small portions into stainless steel pallets pre-cooled and filled with liquid nitrogen (T _bale = -196 ° C). The frozen product on the pallets is transferred to a freeze-drying chamber and freeze-dried at temperatures: -45 ° С… -50 ° С on a condenser; on the product -196 ° C ... + 30 ° C and a residual pressure in the chamber (9-5) 10 ^-2 Torr for 25 hours. The product of freeze-drying is a light, fluffy white powder, placed in a 1000 ml heat-resistant chemical glass , bring it into a drying oven preheated to 80 ° C, kept at this temperature for 1.5 hours and cooled to room temperature for 25.0 hours (cooling rate - 2.2 degrees / hour). The product yield was 14.80 g (98.7 wt.%). According to XRD data, the obtained substance is characterized by a typical diffractogram shown in Fig. 4 and a set of interplanar distances and their intensities that coincide with the corresponding values for the new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is presented in Table. 3. According to thermoanalytical studies, the resulting substance is characterized by a typical differential scanning calorimetry curve presented in Fig. 6, which characterizes a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluorop-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate. The temperatures of the endothermic peaks of the obtained powder are (39.5-48.3) ° C and (120.5-125.7) ° C. According to HPLC, the purity of the new, previously unknown, weakly crystallized β-modification of sofosbuvir is 99.97%.

Example 5. Pharmaceutical composition. For the preparation of 10.00 g of a known composition, including as an active substance a crystalline modification of sofosbuvir and excipients in a ratio (wt.%):

Active substance sofosbuvir 33.33 Mannitol 30.00 Microcrystalline cellulose 29.67 Croscarmellose sodium 5.00 Colloidal silicon dioxide 0.50 Magnesium stearate 1.50

3.33 g of a powder of a new, previously unknown, slightly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxco3,4- dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is mixed for 10 minutes in a ceramic mortar with auxiliary substances: 3.00 g mannitol (according to the US Pharmacopoeia and the European Pharmacopoeia), 2.97 g of microcrystalline cellulose (according to the US Pharmacopoeia and the European Pharmacopoeia), 0.50 g of croscarmellose sodium (according to the US Pharmacopoeia and the European Pharmacopoeia), 0.05 g of silicon wild colloid type (according to the US Pharmacopoeia and the European Pharmacopoeia) and 0.15 g of magnesium stearate (according to TU 6-09-16-1533-90). The resulting mixture was subjected to physico-chemical and biological methods of analysis. According to XRD data, a substance is present in the resulting mixture, which is characterized by a set of interplanar distances and their intensities, which coincide with the corresponding values for the new, not previously known, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy ) phosphorylamino) propanoate, presented in table. 3

In a similar way and of identical composition, a composition was prepared based on the known crystalline modification of sofosbuvir (Form 1), which was used as the starting material. According to XRD data, a substance is present in the resulting mixture, which is characterized by a set of interplanar distances and their intensities, which coincide with the corresponding values for the known modification (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5 - (2,4-dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate (Form 1) presented in table. 2

Biological experiments were performed on male rabbits weighing 2.0-2.5 kg. To obtain reliable results, parallel series of experiments using not less than 5 ^five rabbits. Determination of the time of occurrence of sofosbuvir in the blood was performed by the in vivo method when the composition was orally administered, followed by collection of plasma from the auricle. In all cases, the amount of active substance introduced into the animal was 37.50 mg of the mixture (12.38 mg of sofosbuvir). It turned out that for a composition containing a new, not previously known, crystalline β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo- 3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate in the blood of rabbits is (14 ± 4) min, and for a known composition containing as an active substance a known crystalline modification of sofosbuvir (Form 1) - (25 ± 4) min.

Thus, we have discovered a new, previously unknown, weakly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3, 4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, a method for its preparation and its use for the preparation of a pharmaceutical composition as nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C.

The newly obtained, previously unknown, weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is characterized by a set of interplanar distances (d, Å) different from the known crystalline modifications of sofosbuvir, and the corresponding their intensities (I _rel ,%):

25.638 Å - 15.95%; 21.101 Å - 17.80%; 16.458 Å - 21.57%; 13.368 Å - 26.61%; 12.290 Å - 28.21%; 11.343 Å - 31.14%; 11.032 Å - 40.14%; 01.287 Å - 36.18%; 9.083 Å - 39.89%; 8.458 Å - 43.59%; 7.641 Å - 40.40%; 7.148 Å - 46.78%; 7.013 Å - 54.75%; 6.528 Å - 46.01%; 5.975 Å - 55.33%; 5.225 Å - 79.00%; 5.093 Å - 100.00%; 4.892 Å - 86.15%; 4.707 Å - 98.40%; 4.614 Å - 84.56%; 4.535 Å - 94.38%; 4.505 Å - 90.68%; 4.371 Å - 89.60%; 4.259 Å - 75.49%; 4.008 Å - 62.48%; 3.792 Å - 57.69%; 3.741 Å - 54.50%; 3.539 Å - 47.35%; 3,500 Å - 46.01%; 3.263 Å - 46.27%; 3.172 Å - 38.54%; 3.141 Å - 40.71%; 2.99 Å - 25.82%; 2.839 Å - 25.82%; 2.773 Å - 25.06%; 2.722 Å - 27.38%; 2.522 Å - 25.02%; 2.359 Å - 23.68%; 2.192 Å - 22.85%; 2.117 Å - 21.83%,

and the presence of two endothermic effects on the DSC curve at temperatures (39.7-48.1) ° C and (120.7-125.9) ° C.

In addition, a new, not previously known, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4- dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate is distinguished by a higher biological activity as a part of the pharmaceutical composition compared to the known crystalline modification of sofosbuvir (Form 1), which was used as the starting material.

From the above, we can conclude that the claimed new, not previously known, slightly crystallized β - modification of (S) -isopropyl 2 - ((S) - ((2R, 3R, 4R, 5R) -5- (2.4 -dioxo-3,4-dihydropyrimidine- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, its preparation and its use for the preparation pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C are new and meet the criteria of "inventive step" and "industrial I'm applicability. "

¹⁾ WO 2010/135569 A1 dated November 25, 2010

²⁾ WO 2015/025853 A1 dated March 5, 2015; ³⁾ US 2015/0175646 dated June 25, 2015; ⁴⁾ CN 104125252 A dated November 5, 2014;

⁵⁾ WO 2016/035006 A1 dated 03/10/2016; ⁶⁾ WO 2015/126995 A1 dated 08.27.2015

⁶⁾ WO 2015/126995 A1 dated August 27, 2015; ⁷⁾ WO 2015/191945 A2 dated December 17, 2015; ⁸⁾ WO 2016/038542 A2 dated 03/17/2016; ⁹⁾ WO 2016/008461 A1 dated 01/21/2016

¹⁰⁾ CN 104277088 A dated January 14, 2015; ¹¹⁾ WO 2016/023906 A1 02.18.2016

Claims (6)

1. Weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl ) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate, characterized by the following set of interplanar distances (d, Å) and corresponding intensities (I _rel ,%): 25.638 Å - 15.95%; 21.101 Å - 17.80%; 16.458 Å - 21.57%; 13.368 Å - 26.61%; 12.290 Å - 28.21%; 11.343 Å - 31.14%; 11.032 Å - 40.14%; 01.287 Å - 36.18%; 9.083 Å - 39.89%; 8.458 Å - 43.59%; 7.641 Å - 40.40%; 7.148 Å - 46.78%; 7.013 Å - 54.75%; 6.528 Å - 46.01%; 5.975 Å - 55.33%; 5.225 Å - 79.00%; 5.093 Å - 100.00%; 4.892 Å - 86.15%; 4.707 Å - 98.40%; 4.614 Å - 84.56%; 4.535 Å - 94.38%; 4.505 Å - 90.68%; 4.371 Å - 89.60%; 4.259 Å - 75.49%; 4.008 Å - 62.48%; 3.792 Å - 57.69%; 3.741 Å - 54.50%; 3.539 Å - 47.35%; 3,500 Å - 46.01%; 3.263 Å - 46.27%; 3.172 Å - 38.54%; 3.141 Å - 40.71%; 2.99 Å - 25.82%; 2.839 Å - 25.82%; 2.773 Å - 25.06%; 2.722 Å - 27.38%; 2.522 Å - 25.02%; 2.359 Å - 23.68%; 2.192 Å - 22.85%; 2.117 Å - 21.83%, and the presence of two endothermic effects on the DSC curve at temperatures of 39.7-48.1 ° C and 120.7-125.9 ° C. 2. A method of obtaining a weakly crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) -yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate according to claim 1, characterized in that sofosbuvir is dissolved in an organic solvent at 25-70 ° C, mix the resulting solution with distilled water in a ratio of 1: 2-1: 10, freeze at a cooling rate not lower than 60 deg / min, freeze-drying and additionally anneal at temperatures of 50-85 ° C for at least 1 h with eduyuschim cooled to room temperature at a rate no higher than 5 ° / hr. 3. The method according to p. 2, characterized in that the freeze-drying of the frozen solution of sofosbuvir is carried out at temperatures: on the condenser (-43) - (- 56) ° C; on the product (-196) - (+ 50) ° C and the residual pressure in the chamber (9-3) × 10 ^-2 Topp for 22-26 hours 4. Application of weakly-crystallized β-modification of (S) -isopropyl 2 - ((S) - (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin- (2H) - yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) - (phenoxy) phosphorylamino) propanoate according to claim 1 for preparing a pharmaceutical composition as a nucleotide polymerase inhibitor for the treatment of chronic viral hepatitis C.

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